The Role of Polymer Nanolayer Architecture on the Separation Performance of Anion-Exchange Membrane Adsorbers: Part II. DNA and Virus Separations

被引:13
|
作者
Bhut, Bharat V. [1 ,2 ]
Weaver, Justin [3 ]
Carter, Andrew R. [1 ,2 ]
Wickramasinghe, S. Ranil [3 ]
Husson, Scott M. [1 ,2 ]
机构
[1] Clemson Univ, Dept Chem & Biomol Engn, Clemson, SC 29634 USA
[2] Clemson Univ, Ctr Adv Engn Fibers & Films, Clemson, SC 29634 USA
[3] Colorado State Univ, Dept Chem & Biol Engn, Ft Collins, CO 80523 USA
关键词
ATRP; chromatography; downstream processing; DNA purification; virus clearance; SUPERCOILED PLASMID DNA; HIGH-CAPACITY; CHROMATOGRAPHY; PURIFICATION; ULTRAFILTRATION; TRANSMISSION; SIZE;
D O I
10.1002/bit.23222
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
The surface-initiated polymerization protocol developed in part I was used to prepare strong anion-exchange membranes with variable polymer chain graft densities and degrees of polymerization for DNA and virus particle separations. A focus of part II was to evaluate the role of polymer nanolayer architecture on DNA and virus binding. Salmon sperm-DNA (SS-DNA) was used as model nucleic acid to measure the dynamic-binding capacities at 10% breakthrough. The dynamic-binding capacity increases linearly with increasing poly ([2-(methacryloyloxy)ethyl]trimethylammonium chloride) chain density up to the highest chain density used in this study. The new membranes yielded threefold higher SS-DNA-binding capacity (30 mg/mL) than a leading commercial membrane with the same functional group chemistry. Elution of bound DNA yielded a sharp peak, and resulted in a 13-fold increase relative to the feed concentration. This concentration effect further demonstrates the highly favorable transport properties of the newly designed Q-type membranes. However, unlike findings in part I on protein binding, SS-DNA binding was not fully reversible. Minute virus of mice (MVM) was used as model virus to evaluate the virus clearance performance of newly designed Q-type membranes. Log reduction of virus (LRV) of MVM increased with increasing polymer chain density. Membranes exhibited >4.5 LRV for the given MVM impurity load and may be capable of higher LRV values, as the MVM concentration in the flow-through fraction of these samples was below the limit of detection of the assay. Biotechnol. Bioeng. 2011; 108: 2654-2660. (C) 2011 Wiley Periodicals, Inc.
引用
收藏
页码:2654 / 2660
页数:7
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